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EpiJET 5-hmC Analysis Kit

Kód produktu: K1481 Kód výrobce: K1481 Kód dodavatele: {9341728F-799F-40F0-A71C-79549F5F2A9C} Výrobce: UAB Fermentas
5 620,45 Kč
4 645,00 Kč bez DPH
25 rxns

EpiJET 5-hmC Analysis Kit is used for 5-hmC analysis and quantification in CCGG sequences.

Zobraz detailní popis

Detailní popis

The Thermo Scientific EpiJET 5-hmC Analysis Kit is an efficient system for detection of 5-hydroxymethylcytosine (5-hmC) within specific DNA loci. The kit takes advantage of the T4 phage β-glucosyltransferase (T4 BGT) ability to specifically modify 5-hmC residues by adding glucose moiety to 5-hmC and the sensitivity of restriction enzyme MspI to glucosylated 5-hmC.

Features

The EpiJET 5-hmC Analysis Kit includes two advanced enzymes: T4 BGT and Epi MspI. The T4 BGT is formulated for highly specific, complete, and fast glucosylation of 5-hmC in 15 min. The Epi MspI is an MspI restriction enzyme specially formulated for efficient use in epigenetic studies. It completely digests genomic DNA in 4 hours when cytosine, 5-methylcytosine, or 5-hydroxymethylcytosine is within its recognition sequence, but its activity is completely blocked when 5-hmC is glucosylated. The analysis involves DNA sample glucosylation and subsequent Epi MspI digestion. Restriction digestion products are then analyzed by qPCR with a primer pair flanking CCGG site of interest (see Figure 1 in Supporting Data). Use the specially designed calculator for calculation of 5-hmC percentage. 

Highlights

  • Complete and specific 5-hmC conversion to glucosylated 5-hmC in 15 min
  • Discrimination of 5-hmC in CCGG sequences by Epi MspI digestion in 4 hours
  • Quantification of 5-hmC percentage in CCGG sequences by qPCR

Applications

  • 5-hmC analysis and quantification in CCGG sequences

Includes

  • T4 β – glucosyltransferase, 5 U/µL
  • 10X Epi Buffer
  • 10X UDP–glucose
  • Epi MspI
  • 5-hmC Control DNA
  • Control primer 1
  • Control primer 2
  • Water, nuclease-free .
  • Detailed Protocol
Hazardous No
Quality Control
  • The kit is functionally tested by spiking human blood genomic DNA with 5-hmC Control DNA. The DNA is glucosylated and digested with the Glc-5-hmC sensitive restriction endonuclease Epi MspI.
  • Glucosylation/digestion levels are assayed by qPCR.
  • The kit passes QC requirements if ΔCq value between “+T4 BGT+Epi MspI” and “-T4 BGT-Epi MspI” samples is ≤ 0.5 cycle, and ΔCq value between “-T4 BGT+Epi MspI” and “-T4 BGT-Epi MspI” samples is ≥ 4.5 cycles.
REviewer Use for DNA sequence analysis and map creation.
Storage Condition -20 C

Obsah balení

25 rxns

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